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Wizard miniprep protocol

Steve Stelman steves at burn.ucsd.edu
Tue Dec 12 20:15:59 EST 1995

Surasakdi Wongratanacheewin wrote:

> Does anyone there have the protocol for Wizard miniprep (Promega).
> I have the kit but I loss my protocol. Could you please send me that
> protocol.
>         Thanks for help.
> Surasak

        1) Pellet 1-3 ml cells.
           Resuspend in 200 ul Cell Resuspension Soln. (I)
           Add 200 ul Cell Lysis Soln. (II)
           Invert until soln clears.
           Add 200 ul Neutralization Soln. (III)
           Decant sup.
        2) Add 1 ml Purification Resin.
           Attach 5 ml syringe to column.  Remove plunger.
           Load sample into syringe.
           Gently push into column.
           Detach column, remove plunger, replace column.
        3) Load 2 ml Column Wash Soln.
           Gently push into column.
           Remove column to epitube.
           Spin dry 2 min.
        4) Remove column to clean epitube.
           Load 50 ul TE buffer.
           Wait 1 min.
           Spin 2 min. to elute.
        (I) Cell Resuspension Soln:
                   50 mM  Tris-HCl, pH 7.5
                   10 mM  EDTA
                100ug/ml  RNase A

        (II) Cell Lysis Soln:
                  0.2 M  NaOH
                     1%  SDS

        (III) Neutralization Soln:
                 2.55 M  KOAc, pH4.8

        Column Wash Soln:
                  200mM  NaCl
                   20mM  Tris-HCl, pH 7.5
                    5mM  EDTA
               **dilute 1:1 with 95% EtOH  

PS.  If you want the recipe for the resin (or a working facsimile),
I have a recipe from an old post to this group for "Merlin" resin.

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